ࡅ࠻ࠛࠬ࠻ࡠࠥࡦฃኈ૕⚿วߦࠃࠆᵴᕈൻ࡮ᜟ᛫૞↪‛⾰ࠍᬌ಴ߔࠆ BG1Luc ER TA ᴺߩ⹏ଔળ⼏ႎ๔ᦠJaCVAM ⹏ଔળ⼏ᐔᚑ 25 ᐕ 6 ᦬ 11 ᣣ11JaCVAM ⹏ଔળ⼏ศ↰ᱞ⟤㧔ᣣᧄᲥᕈቇળ㧕㧦ᐳ㐳ᵻ㊁ື⑲㧔ᣣᧄⅣႺᄌ⇣ේቇળ㧕੖ච፲⦟᣿ 㧔࿖┙ක⮎ຠ㘩ຠⴡ↢⎇ⓥᚲ ↢ᵴⴡ↢ൻቇㇱ㧕৻㝩 ീ㧔ᣣᧄൻቇᎿᬺදળ㧕*ᄢፉஜᐘ㧔ᣣᧄൻቇᎿᬺදળ㧕ᄢ㊁ᵏ㓶㧔ᐳ㐳ផ⮈㧕*ዊ═ේᒄ㆏ 㧔⁛┙ⴕ᡽ᴺੱ ක⮎ຠක≮ᯏེ✚วᯏ᭴㧕
ዊ㊁ኹඳᔒ 㧔⁛┙ⴕ᡽ᴺੱ ක⮎ຠක≮ᯏེ✚วᯏ᭴㧕㤥Ỉ ദ㧔ᣣᧄേ‛ታ㛎ઍᦧᴺቇળ㧕᧖ጊ⌀ℂሶ 㧔ᣣᧄൻ♆ຠᎿᬺㅪวળ㧕⼱↰ᥓሶ㧔⁛┙ⴕ᡽ᴺੱ ක⮎ຠක≮ᯏེ✚วᯏ᭴㧕*⷏Ꮉ⑺૫㧔࿖┙ක⮎ຠ㘩ຠⴡ↢⎇ⓥᚲ ቟ోᕈ↢‛⹜㛎⎇ⓥ࠮ࡦ࠲࡯㧕㐳⼱Ꮉ㓉৻ 㧔⁛┙ⴕ᡽ᴺੱ ⵾ຠ⹏ଔᛛⴚၮ⋚ᯏ᭴㧕’ ᩕੑ㧔ᣣᧄ఺∉Ქᕈቇળ㧕Ⴧ↰శノ㧔ᐳ㐳ផ⮈㧕ጊ↰㓉ᔒ㧔⁛┙ⴕ᡽ᴺੱ ⵾ຠ⹏ଔᛛⴚၮ⋚ᯏ᭴㧕*ᮮ㑐ඳ㓶㧔ᣣᧄ⊹⤏ࠕ࡟࡞ࠡ࡯࡮ធ⸅⊹⤏Ἳቇળ㧕ศ↰ ✛㧔࿖┙ක⮎ຠ㘩ຠⴡ↢⎇ⓥᚲ ቟ోᕈ↢‛⹜㛎⎇ⓥ࠮ࡦ࠲࡯ ∛ℂㇱ㧕ศ᧛ ഞ㧔ᐳ㐳ផ⮈㧕ᷰㇱ৻ੱ㧔ᣣᧄ⵾⮎Ꮏᬺදળ㧕                  છᦼ㧦ᐔᚑ 24 ᐕ 4 ᦬ 1 ᣣ㨪ᐔᚑ 26 ᐕ 3 ᦬ 31 ᣣ*㧦ᐔᚑ 25 ᐕ 4 ᦬ 1 ᣣ㨪ᐔᚑ 26 ᐕ 3 ᦬ 31 ᣣએ਄22ౝಽᴲᡬੂ‛⾰߇ᄢ߈ߥ໧㗴ߣߥߞߡ߆ࠄ‫ࡦࠥࡠ࠻ࠬࠛߩ⾰‛ޔ‬᭽૞↪ࠍ᷹ቯߔࠆ in vitro ⹜㛎ᴺߩ㐿⊒߇␠ળ⊛ߦ᳞߼ࠄࠇߡ߈ߚ‫ޕ‬BG1LucER TA ᴺ㧔એਅ‫ޟ‬ᒰ⹥⹜㛎ᴺ‫ޠ‬㧕ߪ‫␠ߩߎޔ‬ળ⊛ⷐ⺧ߦᔕ߃ߡ㐿⊒ߐࠇߚ⹜㛎ᴺߩ৻ߟߢ‫ޔ‬ൻቇ‛⾰߇ ࡅ࠻ࠛࠬ࠻ࡠࠥࡦฃኈ૕㧔ER㧕ߦኻߒߡᵴ
ᕈ‛⾰㧔agonist㧕޽ࠆ޿ߪᜟ᛫ᕈ‛⾰㧔antagonist㧕ߢ޽ࠆߎߣࠍᬌ಴ߒߡ‫ࡦࠥࡠ࠻ࠬࠛޔ‬ᵴᕈ޽ࠆ޿ߪᜟ᛫૞↪ࠍ␜ߔౝಽᴲᡬੂᕈߩ੍᷹ߦᓎ┙ߟ߽ߩߣߒߡឭ᩺ߐࠇߚ߽ߩߢ޽ࠆ 1)‫ޕ‬ᒰ⹥⹜㛎ᴺߦኻߒߡᓥ᧪⹜㛎ᴺߣ⸒߃ࠆ߽ߩߪ⏕┙ߒߡ޿ߥ޿‫ޔߒ߆ߒޕ‬ౝಽᴲᡬੂᕈࠍⷙቯߔࠆᲥᕈߥ޿ߒ↢‛ᵴᕈߦߟ޿ߡߪ߹ߛචಽߦߪ․ቯߐࠇߡ޿ߥ޿ㇱಽ߇ᱷߐࠇߡ߅ࠅ‫ޔ‬ICCVAM ߩ̌Invitro ER and AR binding and TA reference substances list2)̍ߢߪ‫ⶄޔ‬ᢙߩᩮ᜚ࠍ✚วߒߡౝಽᴲᡬੂᕈ
‛⾰ࠍಽ㘃ߒߡ޿ࠆ‫ޕ‬☨࿖ߢߪ 2012 ᐕߦ ICCVAM ߇ᒰ⹥⹜㛎ᴺߩ⹏ଔ⚿ᨐࠍႎ๔ߒߡ޿ࠆ 1)‫ޕ‬OECD ߦ߅޿ߡ߽2012 ᐕ 10 ᦬ 2 ᣣߦߎࠇࠍࠟࠗ࠼࡜ࠗࡦ TG457 ߦขࠅ౉ࠇߡ޿ࠆ 3)‫ߦࠇߎޕ‬ኻߒߡᣣᧄߢߪ‫ޔ‬
JaCVAM ߩ⹏ଔᆔຬળ߇ᒰ⹥⹜㛎ᴺߩ⹏ଔࠍⴕ޿‫ޔ‬ᐔᚑ 24 ᐕ 11 ᦬ߦ⚿ᨐࠍႎ๔ߒߡ޿ࠆ 4㧕‫ޕ‬એਅߦ␜ߔߩߪ‫ߩߎޔ‬ႎ๔ߦၮߠ޿ߚ JaCVAM ⹏ଔળ⼏ߩ⹏ଔ⚿ᨐߢ޽ࠆ‫ޕ‬
1. ᒰ⹥⹜㛎ᴺߪ‫ߥ߁ࠃߩߤޔ‬ᓥ᧪⹜㛎ᴺࠍઍᦧߔࠆ߽ߩ߆‫ߥ߁ࠃߩߤޔߪߚ߹ޕ‬Ქᕈࠍ⹏ଔ޽
ࠆ޿ߪ੍᷹ߔࠆ߽ߩ߆‫ޕ‬ൻቇ‛⾰ߩࠛࠬ࠻ࡠࠥࡦᵴᕈࠍᬌ಴ߒࠃ߁ߣߔࠆ in vivo ߩᓥ᧪⹜㛎ᴺߣߒߡߪ‫ߞߍޔ‬ᱤ㘃ࠍ↪޿ߚሶች⢈ᄢ⹜㛎߇޽ࠆ‫ޕ‬ൻቇ‛⾰ߩࠛࠬ࠻ࡠࠥࡦᵴᕈࠍᬌ಴ߒࠃ߁ߣߔࠆ in vitro ߩ⹜㛎ᴺ
ߣߒߡߪ‫ࡦࠥࡠ࠻ࠬࠛޔ‬ฃኈ૕⚿วᕈ⹜㛎ߣ‫ࡦࠥࡠ࠻ࠬࠛ࠻ࡅޔ‬-Į ฃኈ૕቟ቯዉ౉⚦⢩ࠍ↪޿ࠆォ౮ᵴᕈൻ⹜㛎 (Stably Transfected Human Estrogen Receptor-Į Transcriptional Activation Assay; ER-ĮSTTA; OECD TG455㧕߇޽ࠆ‫ޕ‬ᒰ⹥⹜㛎ᴺߪߎࠇࠄߣ㘃ૃߩᓎഀ߇ᦼᓙߐࠇߡ޿ࠆ߽ߩߢ޽ࠆ‫ޕ‬
2. ᒰ⹥⹜㛎ᴺߣᓥ᧪⹜㛎ᴺߩ㑆ߦߤߩࠃ߁ߥ⑼ቇ⊛ߟߥ߇ࠅ߇޽ࠆ߆‫ޕ‬᷹ቯᜰᮡߪ⇣ߥࠆ߇‫ࡦࠥࡠ࠻ࠬࠛޔ‬ᵴᕈ߹ߚߪᜟ᛫ᕈࠍၮ␆ߣߔࠆߣ޿߁ᗧ๧ߢ‫ޔ‬ᒰ⹥⹜㛎ᴺ
ߩ⑼ቇ⊛ࡔࠞ࠾࠭ࡓߪ‫ߞߍޔ‬ᱤ㘃ሶች⢈ᄢ⹜㛎ߣ౒ㅢߢ޽ࠆ‫ޕ‬㘃ૃߩ⹜㛎ᴺߢ޽ࠆ ER-Į STTA ᴺߪ‫ޔ‬ᵴᕈ‛⾰߇ ER ߣ⚿วߔࠆߎߣߦࠃߞߡ⺃ዉߐࠇࠆDNA ォ౮ᵴᕈൻࠍ࡟ࡐ࡯࠲࡯ㆮવሶ㧔Luciferase㧕ߩᵴᕈൻߦࠃࠆ luciferin ߩ⊒శࠍᜰᮡߣߒߡ᷹ቯߔࠆ߽ߩߢ޽ࠅ‫ޔ‬ᒰ⹥⹜㛎ᴺߩේℂߪߎࠇߣ౒ㅢߢ޽ࠆ‫ޕ‬㘃ૃߩ⹜㛎ᴺߢ޽ࠆࠛࠬ࠻ࡠࠥࡦฃኈ૕⚿วᕈ⹜㛎ߪ‫ⵍޔ‬㛎‛⾰ߣ ER ߣߩ⚿วᕈࠍ᷹ቯߔࠆ߽ߩߢ޽ࠆ߇‫⚿ޔ‬วߒߚ‛⾰߇ ER ࠍೝỗߔࠆߩ߆࡝ࠟࡦ࠼ߦᜟ᛫ߔࠆߩ߆ߪ್೎ߢ߈ߥ޿ߣߎ
ࠈ߇ᒰ⹥⹜㛎ᴺߣ⇣ߥߞߡ޿ࠆ‫ޕ‬
3. ᒰ⹥⹜㛎ᴺߣߘߩ࠺࡯࠲ߪ‫ޔ‬ㅘ᣿ߢ⁛┙ߥ⑼ቇ⊛⹏ଔࠍฃߌߡ޿ࠆ߆‫ޕ‬EU㧔ECVAM㧕ߣᣣᧄ㧔JaCVAM㧕ࠍ૬ߖߡ ICCVAM ߇⚵❱ߒߚㆇ༡ᆔຬળߩ▤ℂߩਅߢ‫ޔ‬3⹜㛎ᣉ⸳㧔☨࿖ߩ Xenobiotic Detection Systems, Inc.‫ޔ‬ECVAM Joint Research Centre‫ޔ‬ᣣᧄߩᣣศᩣᑼળ␠㧕ߦࠃࠆᬌ⸽⹜㛎߇ታᣉߐࠇߚ‫ⵍޕ‬㛎‛⾰ߪ‫ޔ‬ICCVAM ߇ࠛࠬ࠻ࡠࠥࡦ߅ࠃ߮ࠕࡦ࠼ࡠࠥࡦߦ㑐ߔࠆ in vitro ⹜㛎ᴺߩ⹏ଔߦ↪޿ࠆߴ߈‛⾰ߣߒߡ 2003 ᐕߦ⊒⴫ߒߚ 78 ‛⾰ߢ޽ࠆ 2)‫ޕ‬್ቯߦᔅⷐߥ↪㊂෻ᔕᦛ✢߇ᓧࠄࠇߕ‫ࡦࠥࡠ࠻ࠬࠛޔ‬ᵴᕈ࡮ᜟ᛫ᕈߩ᦭ή߇್ቯ࿎㔍ߢ޽ߞߚ 7‛⾰߇‫ߩߎޔ‬ᬌ⸽⹜㛎ߩ⸃ᨆ࡮⹏ଔ߆ࠄ㒰ᄖߐࠇߚߎߣߪ‫ޔ‬ᒰ⹥⹜㛎ᴺߩ೑↪ߦᒰߚߞߡ⇐ᗧߔߴ߈ߎߣߢ޽ࠆ‫ޕ‬33ߎߩ⹜㛎ߦ߅ߌࠆᒰ⹥⹜㛎ᴺߩᱜ⏕ᕈߪ‫ޔ‬ER ᵴᕈ 35 ‛⾰ߦ߅޿ߡ‫ޔ‬ᗵᐲ߇ 96%㧔27/28㧕‫․ޔ‬⇣ᐲ߇ 100%㧔7/7㧕ߢ‫ ߪ₸⥌৻ޔ‬97%㧔34/35㧕ߢ޽ߞߚ‫ޕ‬ER ᜟ᛫ᕈ 25 ‛⾰ߦ߅޿ߡߪ‫ޔ‬ᗵᐲ
߇ 100%㧔3/3㧕‫⇣․ޔ‬ᐲ߽ 100%㧔22/22㧕ߢ‫ ߪ₸⥌৻ޔ‬100%㧔25/25㧕 ߢ޽ߞߚ‫ޕ‬ߎߩ⹜㛎⚿ᨐߦߟ޿ߡߩ࿖㓙⊛╙ਃ⠪⹏ଔߦ߅޿ߡ‫ޔ‬ᒰ⹥⹜㛎ᴺߪ‫ޔ‬ER ᵴᕈ߅ࠃ߮ ER ᜟ᛫ᕈࠍᬌ಴ߔࠆ in vitro ⹜㛎ᴺߣߒߡㆡᱜߥ߽ߩߢ޽ࠆߣߐࠇߡ޿ࠆ‫ޕ‬ࠃߞߡ‫ޔ‬ᒰ⹥⹜㛎ᴺߣߘߩ࠺࡯࠲ߪㅘ᣿ߢ⁛┙ߥ⑼ቇ⊛⹏ଔࠍฃߌߡ޿ࠆߣ⸒߃ࠆ‫ޕ‬
4. ᒰ⹥⹜㛎ᴺߪ‫ޔ‬ᓥ᧪⹜㛎ᴺߩઍᦧᴺߣߒߡ‫⵾ߪߚ߹⾰‛ߥ߁ࠃߩߤޔ‬ຠࠍ⹏ଔߔࠆߎߣࠍ⋡⊛ߣߒߡ޿ࠆ߆‫ޕ‬ઍᦧߔߴ߈⹜㛎ᴺߪ⏕┙ߐࠇߡ޿ߥ޿߇‫ޔ‬ᒰ⹥⹜㛎ᴺߩ⹏ଔኻ⽎ߣߥࠆߩߪ‫ޔ‬ක⮎ຠ‫ޔ‬േ‛↪
ක⮎ຠ‫ޔ‬ㄘ⮎‫↥ޔ‬ᬺൻቇ‛⾰╬ߢ޽ࠆ‫ޔߒߛߚޕ‬ਇṁᕈ‛⾰߅ࠃ߮ើ⊒ᕈ‛⾰ߦኻᔕߔࠆ⹜㛎ᚻ㗅ߪ⏕┙ߐࠇߡ޿ߥ޿‫ޕ‬
5. ᒰ⹥⹜㛎ᴺߪ‫⹏࠼࡯ࠩࡂޔ‬ଔ޽ࠆ޿ߪ࡝ࠬࠢ⹏ଔߩߤߜࠄߦ᦭↪ߢ޽ࠆ߆‫ޕ‬ᒰ⹥⹜㛎ᴺߪ‫ᤨ⃻ޔ‬ὐߢߪࡂࠩ࡯࠼⹏ଔߦ᦭↪ߢ޽ࠆ‫ޕ‬ߥ߅‫ޔ‬ᒰ⹥⹜㛎ᴺߪ‫ⵍޔ‬㛎‛⾰ߩỚᐲߦᔕߓߚᵴᕈߩ᷹ቯ߇น⢻ߢ޽ࠆ߆ࠄ‫⹏ࠢࠬ࡝ޔ‬ଔߦ૶↪ߢ߈ࠆน⢻ᕈߪ޽ࠆ߇‫ߛ߹ޔ‬ᬌ⸽߇ⴕࠊࠇߡ޿ߥ޿ߩߢ‫↪᦭ޔ‬ᕈߪਇ᣿ߢ޽ࠆ‫ޕ‬
6. ᒰ⹥⹜㛎ᴺߪ‫⵾ߪߚ߹⾰‛ࠆߔߣ⊛⋡ޔ‬ຠߩᲥᕈࠍ⹏ଔߢ߈ࠆ߆‫ߩߘޕ‬႐ว‫ޔ‬ᒰ⹥⹜㛎ᴺߩㆡ↪᧦ઙ߇᣿⏕ߦߥߞߡ޿ࠆ߆‫ޕ‬ᒰ⹥⹜㛎ᴺߪ‫ޔ‬ൻቇ‛⾰ߩ ER ᵴᕈ߅ࠃ߮ ER ᜟ᛫ᕈࠍ⺞ߴࠆߎߣ߇ߢ߈ࠆ‫ޕ‬ᒰ⹥⹜㛎ᴺߪ‫ޔ‬ၮᧄ⊛ߥࡊࡠ࠻ࠦ࡞ߣߒߡ‫⹜ޔ‬㛎‛⾰ࠍ DMSO ߦṁ⸃ߒߡㆡ↪ߒߡ߅ࠅ‫ޔ‬DMSO ߦਇṁߩ‛⾰߅ࠃ߮ើ⊒ᕈ‛⾰ߦㆡ↪ߢ߈ࠆ߆ߤ߁߆ߪ‫ޔ‬ᬌ⸛߇ߥߐࠇߡ޿ߥ޿ߩߢਇ᣿ߢ޽ࠆ‫ޕ‬
7. ᒰ⹥⹜㛎ᴺߪࡊࡠ࠻ࠦ࡞ߩᓸ⚦ߥᄌᦝߦኻߒߡ㗎ஜߢ޽ࠆ߆‫ޕ‬ᬌ⸽⹜㛎ߦෳടߒߚ 3 ᣉ⸳ߢߩᣉ⸳ౝౣ⃻ᕈߪ 12 ⵍ㛎‛⾰ߦߟ޿ߡ 100%ߢ޽ߞߚ‫ޕ‬ER ᵴᕈ‛⾰ߦ߅ߌࠆᣉ⸳㑆ౣ⃻ᕈߪ 67%㧔8/12㧕‫ޔ‬ER ᜟ᛫ᕈ‛⾰ߢߩᣉ⸳㑆ౣ⃻ᕈ߽ 100%㧔12/12㧕ߢ޽ߞߚߩߢ‫ࠆ޽ޔ‬⒟ᐲߩ㗎ஜᕈߪᦼᓙߢ߈ࠆ‫ޕ‬8. ᒰ⹥⹜㛎ᴺߩᛛⴚ⠌ᓧߪ‫ޔ‬ㆡಾߥ⸠✵ࠍ⚻ߡ޿ࠆᜂᒰ⠪ߦߣߞߡኈᤃߥ߽ߩߢ޽ࠆ߆‫ޕ‬ߎߩ⹜㛎ߢᔅⷐߥᛛⴚߪ‫ޔ‬ၭ㙃⚦⢩ࠍ↪޿ࠆ⹜㛎ᴺ৻⥸ߩᛛⴚ߅ࠃ߮⚦⢩ߩ⊒శࠍ᷹ቯߔࠆᛛⴚߢ޽ࠅ‫ޔ‬ㆡಾߥ⸠✵ߦࠃߞߡኈᤃߦ⠌ᓧߢ߈ࠆ߽ߩߢ޽ࠆ‫ޕ‬
9. ᒰ⹥⹜㛎ᴺߪ‫ޔ‬ᓥ᧪⹜㛎ᴺߣᲧߴߡᤨ㑆⊛⚻⾌⊛ߦఝࠇߡ޿ࠆ߆‫ޕ‬ᒰ⹥⹜㛎ᴺߩߚ߼ߦᔅⷐߥᯏེߪ‫ޔ‬ㅢᏱߩ⚦⢩ၭ㙃ߦⷐߔࠆⵝ⟎ߩ߶߆‫⚦ޔ‬⢩⊒శߩ᷹ቯߦ↪
޿ࠆశᐲ⸘ߢ޽ࠅ‫ޔ‬㜞ଔߥ߽ߩߢߪߥ޿‫⹜ޕ‬㛎ߦⷐߔࠆᤨ㑆ߪ‫ޔ‬ಓ⚿଻ሽߐࠇߚ⚦⢩ࠍⲢ⸃ߒߡ
Ⴧᱺߔࠆߩߦ 48㨪72 ᤨ㑆‫⹜ޔ‬㛎↪ၭ࿾ߦ⚦⢩ࠍ᠞⒳ߒߡߐࠄߦ 48㨪72 ᤨ㑆‫⹜ޔ‬㛎‛⾰ߣធ⸅ߐߖߡ Luciferase ᵴᕈൻࠍ⺃ዉߔࠆߩߦ 19㨪24 ᤨ㑆ࠍⷐߔࠆ߇‫ߩߘޔ‬ᓟߪ⚦⢩ࠍⲢ⸃ߐߖߡ⊒శࠍ᷹ቯߒ‫ࠍ࠲࡯࠺ޔ‬㓸⸘⸃ᨆߔࠆߩߺߢ޽ࠆ‫ߩࠄࠇߎޕ‬૞ᬺߦⷐߔࠆᤨ㑆ߪઁߩ in vitro 㘃ૃ⹜㛎ᴺߣᄌࠊࠄߥ޿‫ޕ‬44

10. ᒰ⹥⹜㛎ᴺߪ‫ޔ‬േ‛⑔␩ߩⷰὐ෸߮⑼ቇ⊛⷗࿾߆ࠄ‫⵾ߪߚ߹⾰‛ࠆߔߣ⊛⋡ޔ‬ຠߩᲥᕈࠍ⹏
ଔߔࠆઍᦧᴺߣߒߡ‫ⴕޔ‬᡽਄೑↪ߔࠆߎߣߪน⢻߆‫ޕ‬ᒰ⹥⹜㛎ᴺߪၭ㙃⚦⢩ࠍ↪޿ࠆ in vitro ⹜㛎ᴺߢ޽ࠅ‫ߩઁޔ‬㘃ૃ⹜㛎ᴺߣหߓ⒟ᐲߦ‫ⴕޔ‬᡽਄೑↪น⢻ߢ޽ࠆ‫ޕ‬ෳ⠨ᢥ₂1)ICCVAM Test Method Evaluation Report. The LUMI-CELL® ER (BG1Luc ER TA) Test
Method: An In Vitro Assay for Identifying Human Estrogen Receptor Agonist andAntagonist Activity of Chemicals. (NIH Publication No. 11-7814; 2011)2)

ICCVAM Evaluation of In vitro Test Methods for Detecting Potential Endocrine
Disruptors: Estrogen Receptor and Androgen Receptor Binding and Transcriptional
Activation Assays. (NIH Publication No: 03-4503; 2003)

3)

OECD TG457 : BG1Luc Estrogen Receptor Transactivation Test Method for Identifying
Estrogen Receptor Agonists and Antagonists.

4)(2012)

BG1LucER TA (LUMI-CELL ER) ᴺ㧦in vitro ࡅ࠻ࠛࠬ࠻ࡠࠥࡦฃኈ૕ᵴᕈ‛⾰⹜㛎ᴺߩ⹏ଔႎ๔㧔ᐔᚑ 24 ᐕ 11 ᦬㧕55